PTF1A ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Pancreas Specific Transcription Factor 1a (PTF1a) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Pancreas Specific Transcription Factor 1a (PTF1a) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

BHLH transcription factor p48; bHLHa29; class A basic helix-loop-helix protein 29; class II bHLH protein PTF1A; exocrine pancreas-specific transcription factor p48; p48; p48 DNA-binding subunit of transcription factor PTF1; PACA; PAGEN2; pancreas specific transcription factor, 1a; pancreas transcription factor 1 subunit alpha; Pancreas-specific transcription factor 1a; PTF1-p48.

Transcription factor implicated in the cell fate determination in various organs. Binds to the E-box consensus sequence 5'-CANNTG-3'. Plays a role in early and late pancreas development and differentiation. Important for determining whether cells allocated to the pancreatic buds continue towards pancreatic organogenesis or revert back to duodenal fates. May be involved in the maintenance of exocrine pancreas-specific gene expression including ELA1 and amylase. Required for the formation of pancreatic acinar and ductal cells. Plays an important role in cerebellar development. Directly regulated by FOXN4 and RORC during retinal development, FOXN4-PTF1A pathway plays a central role in directing the differentiation of retinal progenitors towards horizontal and amacrine fates.

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Pancreas Specific Transcription Factor 1a (PTF1a) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Pancreas Specific Transcription Factor 1a (PTF1a) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Pancreas Specific Transcription Factor 1a (PTF1a), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Pancreas Specific Transcription Factor 1a (PTF1a) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery Range (%) Average (%) serum (n=5) 96-104 99 EDTA plasma (n=5) 79-88 83 heparin plasma (n=5) 94-102 99 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 96-104 99 serum (n=5) 96-104 99 EDTA plasma (n=5) 79-88 83 EDTA plasma (n=5) 79-88 83 heparin plasma (n=5) 94-102 99 heparin plasma (n=5) 94-102 99

Component Amount Anti-PTF1A Microplate 96 Wells (12 x 8 Well Strips) PTF1A Lyophilized Standard 2 x 10 ng 100X Biotinylated PTF1A Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-PTF1A Microplate 96 Wells (12 x 8 Well Strips) Anti-PTF1A Microplate 96 Wells (12 x 8 Well Strips) PTF1A Lyophilized Standard 2 x 10 ng PTF1A Lyophilized Standard 2 x 10 ng 100X Biotinylated PTF1A Detector Antibody 120 uL 100X Biotinylated PTF1A Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL

This assay has high sensitivity and excellent specificity for detection of Pancreas Specific Transcription Factor 1a (PTF1a) . No significant cross-reactivity or interference between Pancreas Specific Transcription Factor 1a (PTF1a) and analogues was observed.